Rapid Detection and Identification of Wuchereria bancrofti, Brugia malayi, B. pahangi, and Dirofilaria immitis in Mosquito Vectors and Blood Samples by High Resolution Melting Real-Time PCR.
10.3347/kjp.2013.51.6.645
- Author:
Tongjit THANCHOMNANG
1
;
Pewpan M INTAPAN
;
Chairat TANTRAWATPAN
;
Viraphong LULITANOND
;
Sudchit CHUNGPIVAT
;
Piyanan TAWEETHAVONSAWAT
;
Worasak KAEWKONG
;
Oranuch SANPOOL
;
Penchom JANWAN
;
Wej CHOOCHOTE
;
Wanchai MALEEWONG
Author Information
1. Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. wanch_ma@kku.ac.th
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Wuchereria bancrofti;
Brugia malayi;
Dirofilaria immitis;
Brugia pahangi;
high resolution melting real-time PCR;
dog;
mosquito
- MeSH:
Animals;
Blood/*parasitology;
Brugia/classification/genetics/*isolation & purification;
Cats;
Culicidae/*parasitology;
Dirofilaria immitis/classification/genetics/*isolation & purification;
Dogs;
Humans;
Male;
Parasitology/*methods;
RNA, Helminth/genetics;
RNA, Ribosomal, 5S/genetics;
Real-Time Polymerase Chain Reaction/*methods;
Sensitivity and Specificity;
Transition Temperature;
Wuchereria bancrofti/classification/genetics/*isolation & purification
- From:The Korean Journal of Parasitology
2013;51(6):645-650
- CountryRepublic of Korea
- Language:English
-
Abstract:
A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at 81.5+/-0.2degrees C, 79.0+/-0.3degrees C, 76.8+/-0.1degrees C, and 79.9+/-0.1degrees C, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors.
